A robust method to screen detergents for membrane protein stabilization, revisited

被引：11

作者：

Champeil, Philippe ^{[1
]}

Orlowski, Stephane ^{[1
]}

Babin, Simon ^{[1
]}

Lund, Sten ^{[2
]}

le Maire, Marc ^{[1
]}

Moller, Jesper ^{[3
,4
]}

Lenoir, Guillaume ^{[1
]}

Montigny, Cedric ^{[1
]}

机构：

[1] Univ Paris Saclay, Univ Paris Sud, CNRS, I2BC,CEA, F-91198 Gif Sur Yvette, France

[2] Aarhus Univ Hosp, Dept Endocrinol & Internal Med, Med Res Lab, Aarhus, Denmark

[3] Aarhus Univ, Danish Natl Res Fdn, PUMPKIN, Ctr Membrane Pumps Cells & Dis, DK-8000 Aarhus, Denmark

[4] Aarhus Univ, Dept Biomed, DK-8000 Aarhus, Denmark

来源：

ANALYTICAL BIOCHEMISTRY | 2016年 / 511卷

关键词：

Detergent; Membrane protein; Lipid; Stability; LMNG; SERCA1a; SARCOPLASMIC-RETICULUM; SOLUBILIZATION; CA2+-ATPASE;

D O I：

10.1016/j.ab.2016.07.017

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

This report is a follow up of our previous paper (Lund, Orlowski, de Foresta, Champeil, le Maire and Moller (1989), J Biol Chem 264:4907-4915) showing that solubilization in detergent of a membrane protein may interfere with its long-term stability, and proposing a protocol to reveal the kinetics of such irreversible inactivation. We here clarify the fact that when various detergents are tested for their effects, special attention has of course to be paid to their critical micelle concentration. We also investigate the effects of a few more detergents, some of which have been recently advertised in the literature, and emphasize the role of lipids together with detergents. Among these detergents, lauryl maltose neopentyl glycol (LMNG) exerts a remarkable ability, even higher than that of beta-dodecylmaltoside (DDM), to protect our test enzyme, the paradigmatic P-type ATPase SERCA1a from sarcoplasmic reticulum. Performing such experiments for one's favourite protein probably remains useful in pre-screening assays testing various detergents. (C) 2016 Elsevier Inc. All rights reserved.

引用

页码：31 / 35

页数：5

共 15 条

[1] PERTURBATION OF THE STRUCTURE AND FUNCTION OF A MEMBRANOUS CA-2+-ATPASE BY NON-SOLUBILIZING CONCENTRATIONS OF A NON-IONIC DETERGENT [J].